^3H—TdR掺入法检测鸡不同组织淋巴细胞增殖反应的条件优化

Optimal Conditions of ^3H-TdR for Chicken Lymphocyte Transformation Response in Different Immune Tissues

为了检测感染免疫抑制病毒后鸡群的细胞免疫变化，以建立的^3H—TdR掺入法观察了外周血、脾脏、胸腺、法氏囊中淋巴细胞的增殖反应，对影响该方法的细胞密度、丝裂原浓度（ConA或LPS）、犊牛血清（FCS）浓度等条件进行优化。结果表明，外周血淋巴细胞增殖试验的最佳条件：细胞10^5～10^6／mL，ConA5mg／L，FCS10％；脾脏中淋巴细胞最佳条件：细胞5×10^6／mL，ConA10mg／L，FCS10％；胸腺中淋巴细胞最佳条件：细胞10^6／mL，ConA40mg／L，FCS10％；法氏囊中淋巴细胞最佳条件：细胞5×10^6／mL，LPS40mg／L，FCS10％。

^3H-thymidine （3H-TdR） incorporation assay has been used to measure chicken lymphocyte proliferation in peripheral blood,spleen,thymus and bursa. The effect of various parameters including concentration of lymphocyte,concanavalin A （ConA） or bacterial lipopolysaccharide（LPS） and percentage of heat inactivated fetal calf serum（FCS） incubation time were evaluated. The results showed that,the highest response of peripheral blood T-cells to ConA was 5 mg/L at 105-106/mL in RP- MI1640 including 10% FCS during incubation for 72 h at 37 C ,that of spleen T-cells to ConA was 10 mg/L at 53〈 106/mL in RPMI1640 including 10% FCS,that of thymus T-cells to ConA was 40 mg/L at 106/mL in RPMI1640 including 10% FCS and that of bursa B-cells to LPS was 40 mg/L at 5）〈 106/mL in RPMI1640 including 10% FCS. 3H-TdR incorpgration assay is useful to analyze lymphocyte proliferation of avian infected with different immunosuppression viruses. ;chicken